1. Effects of Intravenous Immunoglobulins on Mice with Experimental Epidermolysis Bullosa Acquisita 
Misa Hirose, Benjamin Tiburzy, Norito Ishii, Elena Pipi, Sabina Wende, Ellen Rentz, Falk Nimmerjahn, Detlef Zillikens, Rudolf A. Manz, Ralf J. Ludwig, Michael Kasperkiewicz 
Journal of Investigative Dermatology 
Volume 135, Issue 3, Pages (March 2015)
DOI: /jid
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2. Figure 1
Intravenous immunoglobulins (IVIG) treatment improves clinical and histological disease in mice with experimental epidermolysis bullosa acquisita (EBA). Clinical scores were calculated as the percentage of the body surface area covered by EBA lesions and normalized to 1 before the start of treatment and after reaching scores of ⩾2%. (a) Mean scores are shown as clinical courses of phosphate-buffered saline (PBS)-, methylprednisolone (MP) (20 mg kg day-1 i.p.)-, and IVIG (2 g kg-1 once per week i.p.)-treated mice during a 4-week treatment period. (b) Calculation of the area under the curve (AUC) revealed that the disease severity of IVIG- but not MP-treated mice was significantly reduced compared with nontreated animals. (c–e) Representative clinical and (f–h) histological presentations from PBS-, MP-, and IVIG-treated mice, respectively, at the end of the 4-week treatment period. (i) Comparison of the dermal neutrophil infiltration between the different treatment groups revealed a significantly lower skin myeloperoxidase (MPO) level in the IVIG-treated mice only. Significances were calculated with ANOVA followed by an all pairwise multiple comparison procedure (Dunn’s method) and expressed as mean±SEM (28–44 and 10–13 mice per group for clinical and MPO analysis, respectively). *P<0.05 (in a, each first and second asterisk corresponds to P<0.05 between PBS-treated mice and IVIG- or MP-injected animals, respectively; ns, not significant). Scale bars=100 μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
IVIG attenuate but do not arrest post-treatment disease progression in EBA mice. The mean extent of skin lesions significantly increased within the 4-week treatment period and a further significant mean increase in disease severity was observed in a subgroup of vehicle-treated mice which were followed for 8 additional weeks following last PBS injection (indicated by P<0.05 in the upper brackets). In contrast, disease progression was suppressed in a subgroup of IVIG-treated animals during the treatment period, and after discontinuation of IVIG therapy clinical disease continuously worsened, but this was not statistically significant (indicated by ns in the lower brackets). Direct comparison of control with IVIG-treated mice revealed significant differences in disease extent for up to 3 weeks after treatment but not any longer by the end of the observation period. Significances were calculated with the Wilcoxon rank sum test and repeated measure ANOVA and expressed as mean±SEM (8–11 mice per group). *P<0.05 (referred to differences between PBS- and IVIG-treated animals).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
IVIG reduce circulating autoantibodies and skin-bound immunoreactants in EBA mice. (a) IVIG- but not MP-treated mice showed significantly lower levels of circulating anti-type VII collagen (COL7)-specific IgG compared with PBS-treated mice at the end of the 4-week treatment period, as shown by ELISA. (b) In contrast, IVIG had no significant impact on the amount of total circulating IgG. (c, d) Significantly lower staining intensity of skin basal membrane-bound IgG and C3 was found in IVIG- but not MP-treated mice compared with PBS-treated mice at the end of the 4-week treatment period, as shown by direct immunofluorescence microscopy. (e, f) Deposition of complement fixing IgG2b/c but not noncomplement-fixing IgG1 at the dermal–epidermal junction was significantly reduced in IVIG- compared with PBS-treated mice at the end of the 4-week treatment period, as shown by direct immunofluorescence microscopy. (g) A significantly higher (IgG2b+IgG2c)/IgG1 ratio obtained from skin evaluation was found in the PBS-treated animals compared with IVIG-treated mice. Significances were calculated with ANOVA followed by an all pairwise multiple comparison procedure (Dunn’s method) and expressed as mean±SEM (38–44 mice per group). *P<0.05. Scale bars=50 μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
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5. Figure 4
IVIG decrease FcγRIV expression on circulating Gr-1-positive cells in EBA mice. (a) Significantly lower expression (mean fluorescence intensity, MFI) of FcγRIV on peripheral blood Gr-1-positive cells was observed in IVIG-treated compared with PBS-treated mice after the 4-week treatment period. (b) Representative corresponding flow cytometric analysis. Significances were calculated with a t-test and expressed as mean±SEM (seven mice per group). *P<0.05.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions